Background: The current higher manufacturing cost of biofuels production from lignocellulosics hinders the commercial process development. Although many approaches for reducing the manufacturing cost of cellulosic biofuels may be considered, the use of less expensive feedstocks may represent the largest impact. In the present study, we investigated the use of a low cost feedstock: post-biogas digestion dairy manure fiber. We used an innovative pretreatment procedure that combines dilute sodium hydroxide with supplementary aqueous ammonia, with the goal of releasing fermentable sugar for ethanol fermentation.